KR101357814B1 - The vegetable cultivating method to use fermented liquid chitooligosaccharide fertilizer - Google Patents

Abstract

The present invention relates to a method for cultivating fruit vegetables using chitooligosaccharide fermentation broth, and more specifically, chitosan having a molecular weight of 200,000 or more is fermented to a culture strain of Lyzopus strain and decomposed into oligomers having a molecular weight of 10,000 or less and then irrigated to fruit vegetables with liquid ratio. As a result of foliar fertilization, the present invention relates to a method for cultivating fruit vegetables using chitooligosaccharide fermentation broth, in which the chitooligosaccharide is detected as a fruit vegetable body component in the fruit vegetables, thereby producing fruit vegetables having a specific function.

Description

The vegetable cultivating method to use fermented liquid chitooligosaccharide fertilizer}

The present invention relates to a method for producing fruit vegetables using a chitooligosaccharide fermentation broth ratio, and more particularly, chitosan having a molecular weight of 200,000 or more is fermented to a culture strain of Lyzopus strain and decomposed into oligomeric form having a molecular weight of 10,000 or less and then irrigated to fruit vegetables with liquid ratio. As a result of foliar fertilization, the present invention relates to a method for cultivating fruit vegetables using chitooligosaccharide fermentation broth, in which the chitooligosaccharide is detected as a fruit vegetable body component in the fruit vegetables, thereby producing fruit vegetables having a specific function.

To date, there are few products that absorb specific active ingredients into fruit vegetables to convert and express them into body ingredients, and there are few techniques and cultivation methods for converting chitosan into fruit ingredients by converting chitosan into active ingredients, which is generally known as an excellent polymer. In addition, chitosan has a high molecular weight, making it difficult to absorb melons and low molecular weight studies to absorb them.

In addition, functional agricultural products are released according to the development of the cultivation method, but the verification of the effectiveness of the functionality itself has not been made, but various methods for absorbing conventional chitosan in agricultural products such as melons have been proposed.

Patent No. 0880603 relates to a water treatment plant sludge treatment method using chitosan and chitosan degrading enzyme and a fertilizer manufacturing method using the same. In order to decompose high molecular weight chitosan used in low molecular weight by using chitosan degrading enzyme which is beneficial to plants such as lycopus, and to make sludge produced by this method, landfill or marine sludge generated by general water treatment chemicals It suggests technologies to prevent secondary environmental pollution caused by dumping and to maximize the growth and development effect of chitosan on plants.

Patent No. 0880990 relates to a method for treating stone powder sludge using chitosan and chitosan degrading enzyme, and a method for preparing fertilizer using the same. In order to decompose high molecular weight chitosan used in low molecular weight by using chitosan degrading enzyme which is beneficial to plants such as lycopus, and to make sludge produced by this method, landfill or marine sludge generated by general water treatment chemicals It suggests technologies to prevent secondary environmental pollution caused by dumping and to maximize the growth and development effect of chitosan on plants.

Patent No. 0992213 relates to a method for producing low molecular chitosan using Ryzopus and a method for producing chitosan-containing plants using the same, the method comprising: dissolving chitosan in a lactic acid solution; Mixing low-cost chitosan having a molecular weight of 3600 or less by mixing the culture of Lyzopus oligosporus KCCM 10624 with the chitosan-dissolved solution; And it provides a low-molecular chitosan-containing tomato production method comprising the step of administering the low-molecular chitosan to the tomato.

Each of the above techniques has been proposed to decompose chitosan into low-molecular chitosan, and produce low-molecular chitosan-produced agricultural products by administering the low-molecular chitosan to produce, but it takes a long time to decompose chitosan into low-molecular chito-oligosaccharide form. There were problems that were required and difficult for mass production.

The present invention has been developed to prepare a method for producing low molecular weight chitooligosaccharides under conditions that can be absorbed and contained in the oligomeric form by breaking down the polymer chitosan of the active ingredient in an oligomer form in a significantly shorter reaction time. It is intended to improve the usability of processed products through various commercialization of functional (chito-oligosaccharide-containing) fruits and vegetables by establishing a method for producing fruits and vegetables containing chitooligosaccharides as body components.

In addition, the present invention is to provide a method for cultivating fruit vegetables using the chito oligosaccharide fermentation broth ratio can be produced in a short time by producing a chito oligosaccharide.

In order to achieve the above object, the present invention comprises the steps of mixing a chitosan degrading enzyme culture in an aqueous solution of 0.1 to 3% by weight of chitosan of 200,000 (Mw) or more; Culturing the chitosan degrading enzyme by leaving the mixture of the chitosan degrading enzyme culture and the chitosan aqueous solution for 1 to 4 hours; Filling a stirrer with an aqueous solution of 0.1 to 3% by weight of chitosan having a molecular weight of 200,000 (Mw) or more, and injecting a mixture of the chitosan degrading enzyme into the stirrer; Putting the chitosan degrading enzyme cultured mixture into the stirrer and then stirring it at high speed for 5 to 15 minutes at a speed of 500 to 1500 rpm; Stirring at a low speed for 2 to 10 hours at 100 to 300 rpm after the high speed stirring; It provides a method for cultivating fruit vegetables using chito oligosaccharide fermentation broth ratio comprising the step of administering the chitooligosaccharide produced after completion of the low speed stirring to fruit vegetables.

Chitosan degrading enzyme in the present invention, Rhizopus sp., Streptomyces griseus, Bacillus sp. Trichoderma hamatum, Trichoderma hagianum (Trhchoderma) harzianum), Pseudomonas putida or Pseudomonas aeruginosa strains can be used. Of these, the rhizosphere strain of rhizopus bacteria is very effective in promoting plant growth and increasing disease resistance.

In addition, in the present invention, chitosan can be decomposed by stirring while introducing air into the stirrer during low speed stirring. Chitosan degrading enzymes are aerobic strains, so when agitated while supplying air, air can catalyze and decompose chitosan quickly.

In addition, in the step of administering the chitooligosaccharide in the present invention, by separating the liquid component and the solid component, the liquid component is foliar fertilization and the solid component can be sprayed on the soil. Although it can be used as it is decomposed into chitooligosaccharide, it can also be used to separate the liquid and solid content.

The present invention has the advantage of producing a low molecular weight chito oligosaccharides under conditions that can be absorbed and contained in the oligomeric form by breakdown of oligomeric polymer chitosan of the active ingredient in a significantly short reaction time compared to the above invention.

In addition, the present invention has the advantage that it is possible to commercialize a variety of functional (chito-oligosaccharide-containing) fruits and vegetables containing the chitooligosaccharide as a body component by applying a low molecular weight chitooligosaccharide to fruit vegetables.

In addition, the present invention has the advantage that the mass production is possible by preparing a chitooligosaccharide in a short time.

In addition, unlike conventional chitosan liquid fertilizers that are supplied to the soil by foliar fertilization or spraying methods, the soil and chitosan are uniformly and electrostatically combined to form a grain structure, which is highly breathable and moisturizing. It is possible to improve soil rooting and improve intellect by continually improving the root development of crops and activating beneficial microorganisms in soil without affecting change.

1 is a flow chart of the present invention;
Figure 2 is a photograph of the strain of Raizus according to the invention
Figure 3 is a graph of chitosan viscosity change with or without chitosan degrading enzyme according to the present invention.
Figure 4 is a graph of chitooligosaccharide detection according to the ratio of the lysopus strain and fermentation broth ratio in the state that the decomposition metabolite of grape foil is not added.
5 is a graph of detection of chitooligosaccharides according to the input amount of the metabolite under the same conditions as the ratio of the strain of Lyzopus and chitosan fermentation broth.
Figure 6 is a graph of detection of chitooligosaccharides according to the input amount of the metabolite under the same conditions as the input amount of the fermented liquor ratio of Lyzopus strain.
Figure 7 is a graph of detection of chitooligosaccharides according to the chitosan fermentation broth ratio and metabolites input under the same input conditions of the strain of Raizus.
Figure 8 is a graph of detection of chitooligosaccharides according to the input conditions of the strain of Lyzopus, chitosan fermentation broth and metabolites.

Hereinafter, with reference to the accompanying drawings will be described in more detail the cultivation method of fruit vegetables using the chitooligosaccharide fermentation broth ratio according to an embodiment of the present invention.

As shown in FIG. 1, a method of cultivating fruit vegetables using chitooligosaccharide fermentation broth ratio according to the present invention is first made of an aqueous solution of 0.1 to 3% by weight of chitosan having a molecular weight of 200,000 (Mw) or more. Chitosan degrading enzyme strain culture is mixed with the prepared chitosan aqueous solution (S1). The mixture is prepared by mixing 3 kg of chitosan degrading enzyme strain culture in 3L of 0.1-3% by weight of chitosan aqueous solution, and the powder form enters the aqueous solution to form a dough, which is left in the dough form for 1 to 4 hours to decompose chitosan. The enzyme will be cultured (S2). Chitosan degrading enzyme strain culture includes a strain capable of producing chitose, chitinase, and various substances capable of supplying proteins to the strain.

Next, the stirrer is filled with a predetermined amount of 0.1-3 wt% chitosan aqueous solution having a molecular weight of 200,000 (Mw) or more. Filled chitosan aqueous solution fills 97L. Combine 3 L of chitosan degrading enzyme culture and 97 L of the aqueous solution filled with agitator to 100 L. In the chitosan aqueous solution filled in the stirrer, a mixture of chitosan degrading enzymes is introduced (S3).

Next, after inputting the mixture cultured chitosan degrading enzyme in the stirrer is subjected to a high speed stirring step for 5 to 15 minutes at a speed of 500 ~ 1500 rpm (S4). High-speed stirring is to ensure that chitosan degrading enzyme is mixed evenly in the aqueous chitosan solution. The chitosan digestion can be quickly and sufficiently distributed in chitosan aqueous solution prior to digesting the chitosan.

After the high speed agitation, the stirrer is subjected to a low speed agitation for 2 to 10 hours at 100 to 300 rpm (S5). Low speed agitation is continued at low speed to increase the contact area to allow continuous contact between chitinase and chitosan. While stirring at low speed, air may be added to the stirrer and stirred (S5 '). Since the strain is an aerobic strain, when stirring while injecting air, the air performs a catalytic role to decompose chitosan in a short time.

After the low speed stirring, chitosan is decomposed into chitooligosaccharide, and the chitooligosaccharide is administered to fruit vegetables (S6). Chitooligosaccharide administered to fruit vegetables can be added to the leaves or soil in the form of a liquid or solid or a mixture of solid and liquid.

<Selection of useful microorganisms>

Experimental conditions: Microbial culture and culture conditions for chitosan fermentation

-Temperature: 30 ℃

-Culture humidity: 60%

-Types of microorganisms: Selection of enzymes that can degrade or ferment natural polymer and soil fertilizer components

Mycelial Growth by Type of Root Microorganisms Types of microorganisms Mycelial Growth Rhizopus sp . ) +++++ Streptomyces griseus ) ++ Bacillus sp.) +++ Trichoderma hamatum ) ++ Trichoderma harzianum ) + Pseudomonas putida ) + Pseudomonas Pseudomonas aeruginosa ) ++

The degree of mycelial growth by type of useful microorganisms was the best ( Rhizopus sp. ) And the optical micrograph is shown in FIG.

< Rhizopus sp .On by Chitooligosaccharide ( chitooligosaccharide A) Manufacturing>

3 is a view showing a change in viscosity with time for a sample containing chitosan degrading enzyme and a sample containing chitosan degrading enzyme according to the present invention. As a result of examining the change in viscosity over time, it was confirmed that the difference between the addition and the addition of the enzyme was apparent. It can be seen that the initial viscosity with the enzyme is much lower than without the enzyme. There are two reasons for this. First, if you look at the graph without the enzyme, you can see that the viscosity decreases slightly over time, which means that there is a change over time when chitosan is slightly decomposed by acetic acid. Therefore, when measured with an enzyme-containing sample, the chitosan sample may have a lower viscosity than the sample used in the previous experiment, and it may be possible that the decomposition has already started and the viscosity decreased further during mixing after adding the second enzyme.

  Reasons for unstable viscosity data after the start of the experiment may be that the bubbles contained in the sample were bursting and the stimulus applied by the instrument may have taken some time to become steady. .

As shown in the figure, it can be easily understood that the case with and without the chitosan degrading enzyme is clearly distinguished. When the enzyme is added, the time for the decomposition of chitosan to be steady is about 7200 seconds, that is, about 2 hours to complete the decomposition.

Therefore, it can be seen that by the method according to the present invention, the time to decompose chitosan into chitooligosaccharide is significantly reduced to within 3 hours.

<Administration to fruit vegetables>

Fermented chitooligosaccharides are foliarly applied to fruit vegetables or fertilized to the soil. It can be administered in the form of liquid or air, or it can be administered to fruit vegetables in a form containing liquid and solids together. When fermented chito-oligosaccharide is administered to fruit vegetables, in order to maximize the content of fruit vegetable body components, the fermented chito-oligosaccharides are processed together with the metabolites that decompose grapes and oils into microorganisms and processed in fruit vegetables to maximize the content of chito-oligosaccharides in functional fruit vegetables. can do.

<Selection of Fruits and Vegetables>

Using the chitooligosaccharide melon cultivation system, we commissioned an accredited institution (testing method of fertilizer quality) to verify the crop effect test including the early growth effect of melon. The results of the study on the leaves showed that the organic fertilizer treatment showed the most increase compared to the control, and the leaves increased by 113 ~ 114%, promoting the growth of crops. The crop growth promoting effect was increased by% ~ 108%.

The early stage of growth and the symptoms and physiological disorders were not observed in the melon cultivation by this test procedure. Under the same conditions, Rhizopus sp., Chitooligosaccharide fermentation broth, and grape breakdown metabolites were treated. Other crops, such as paprika, bell peppers and red peppers, were grown in test packaging.

<Farmhouses and Cultivated Plants with Cultivation System> phrase
minute kind Formal day Modified date Estimated Shipping Date
(40-45 days after modification) Treatment history One Rich Honey / Ace Plus 3/10 4/10-4/15 5/20 Treatment of chito-oligosaccharide fermentation solution ratio, grape park metabolite, lycopus strain, etc. 2 Obok / Tarzan 3/6 4/11 Around the end of May 3 Obok / Lava throne 3/12 Early April 5/20 4 Prestigious / Full-Life 2/13 4/10 5/20 5 the great Wall 3/20 4/20 Around the end of May 6 Great Wall / Soldiers 2/30 4/10 Around the end of May 7 Ober, Gold / Tarzan 2/31 End of March Mid-May 8 Obok / Great Wall 3/25 4/26 Around the end of May

<Processing Conditions According to Melon Cultivation Period (Based on 200 pyeong)> division microbe Fermentation liquid ratio Enzyme Remarks One One One 0 unit:
Ryzopus strain: 3 kg
Fermentation broth ratio: 10 L
Metabolite: 5Kg 2 2 One 0 3 3 One 0 4 One 2 0 5 2 2 0 6 3 2 0 7 One One One 8 2 One One 9 3 One One 10 One 2 2 11 2 2 2 12 3 2 2

< Quitooligosaccharides  Detection Analysis>

The produced chitooligosaccharide melon was commissioned by the Korea Food Industry Association to obtain a report on the content of chitooligosaccharide, and the trends of detection of chitooligosaccharide by microbial agent, chitosan fermentation solution ratio and enzyme concentration were as follows.

<Analyzing Trends of Chitooligosaccharides> division Ryzopus strain Fermentation
Liquid script
product February In March April In May June In July Remarks Example a One One 0 - - 0 - 0.08 0.05 unit:
Ryzopus strain: 3 kg
Fermentation broth ratio:
10L
Metabolites:
5Kg Example b 2 One 0 - - - - 0.11 0.10 Example c 3 One 0 - - - - 0.10 0.10 Example d One 2 0 - - 0.16 - 0.31 0.32 Example e 2 2 0 - - 0.17 - 0.30 0.33 Example f 3 2 0 - - 0.17 - 0.35 0.36 Example g One One One - - 0.18 - 0.25 0.22 Example h 2 One One - - 0.19 - 0.25 0.23 Example i 3 One One - - 0.22 - 0.28 0.24 Example j One 2 2 - - 0.28 - 0.50 0.52 Example k 2 2 2 - - 0.34 - 0.52 0.51 Example l 3 2 2 0 0.01 0.37 - 0.58 0.55 Comparative example m 3 0 2 - - 0 - 0 0

When the melon was cultivated with different amounts of Rhizopus sp., Chitosan fermentation broth, and grapeseed metabolite, the effects on the content of chitooligosaccharides detected as the melon body composition were examined.

First, as shown in Comparative Example m of Table 4, chitooligosaccharide was not detected as a body component in the melon grown only by the Lysopus strain and the metabolite treatment, which was effected by the effect of the chitosan fermentation broth ratio invented by the above technique. It was found that oligosaccharides were contained.

Next, Figure 4 is a graph of chitooligosaccharide detection according to the input amount of the Lysopus strain and fermentation broth in the state that the decomposition metabolite of grape foil is not added. The detection amount of chitooligosaccharides according to the input amount of Ryzopus strain and fermentation broth ratio was different according to the cultivation area when compared under the same conditions. There was a tendency to increase.

Next, FIG. 5 and FIG. 6 are graphs of detection of chitooligosaccharides according to the dose of metabolite under the same conditions as the input amount of the fermented liquor ratio of Lyzopus strain and chitosan. The amount of chitooligosaccharide detected by the metabolite was increased under the same conditions, but the metabolite showed a synergistic effect on the detection of chitooligosaccharide.

Next, Figure 7 is a graph of detection of chitooligosaccharides according to the ratio of chitosan fermentation broth ratio and metabolites under the same input conditions of the Lysopus strain. The effects of chitosan fermentation broth ratio and metabolites on the treatment conditions of the same strains of Lyzopus were confirmed to have a greater effect on chitosan fermentation broth ratio than metabolites.

Next, Figure 8 is a graph of the detection of chitooligosaccharides according to the input conditions of the strain of Lyzopus, chitosan fermentation broth and metabolites. As shown in Fig. 4, it was possible to obtain chitooligosaccharide-containing melon even with the ratio of the lycopus strain and the chitosan fermentation broth, but the chitooligosaccharide content was higher when the enzyme was added, as shown in FIGS. 5 and 6. This tendency is clearly seen in FIG. 8. Therefore, it was confirmed that the lyzopus strain and the metabolite maximize the chitooligosaccharide content in the fruit vegetable.

9 to 11 is a test report for the plants administered chitooligosaccharide fermentation broth ratio according to the present invention. 9 is 0.37 (mg / g) of chitooligosaccharide as a test report for melon, FIG. 10 is 0.03 (mg / g) of chitooligosaccharide as a test report for paprika, and FIG. 11 is a test report for chili pepper. It is indicated that the oligosaccharide contains 0.19 (mg / g).

While the invention has been shown and described with reference to certain preferred embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the above description does not limit the scope of the present invention, which is defined by the limitations of the following claims.

Claims (4)

Mixing chitosan degrading enzyme strain culture in an aqueous solution of 0.1 to 3 wt% chitosan having a molecular weight of 200,000 (Mw) or more;
Culturing the chitosan degrading enzyme by leaving the mixture of the chitosan degrading enzyme culture and the chitosan aqueous solution for 1 to 4 hours;
Filling a stirrer with an aqueous solution of 0.1 to 3% by weight of chitosan having a molecular weight of 200,000 (Mw) or more, and injecting a mixture of the chitosan degrading enzyme into the stirrer;
Putting the chitosan degrading enzyme cultured mixture into the stirrer and then stirring it at high speed for 5 to 15 minutes at a speed of 500 to 1500 rpm;
Stirring at a low speed for 2 to 10 hours at 100 to 300 rpm after the high speed stirring;
Administering chitooligosaccharides produced after completion of the low speed stirring to fruit vegetables;
Method of cultivating fruit vegetables using chitooligosaccharide fermentation broth ratio, characterized in that consisting of.
The method according to claim 1,
The chitosan degrading enzyme is Rhizopus sp., Streptomyces griseus, Bacillus sp. Trichoderma hamatum, Trichoderma harzianum. , Pseudomonas putida (Pseudomonas putida) or Pseudomonas aeruginosa (Pseudomonas aeruginosa) any one of the strains, characterized in that the cultivation method of fruit vegetables using chito oligosaccharide fermentation broth.
The method according to claim 1 or 2,
The method of cultivating fruits and vegetables using the chitooligosaccharide fermentation broth, characterized in that the stirring while the air is stirred in the low speed stirring.
The method according to claim 1,
The step of administering the chitooligosaccharide, liquid component and solid content by separating the liquid foliar fertilization and solid content is characterized in that the spraying on the soil, the cultivation method of fruit vegetables using the chitooligosaccharide fermentation broth ratio.

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